Ca2+-permeable AMPA receptors are activated under less restrictive conditions than the NMDA receptor, are found in numerous neuron types, and may regulate pathways that are more generally controlled by NMDAR Ca2+ currents. We have shown that Ca2+-permeable AMPARs, like the NMDAR, can activate neuronal nitric oxide synthase (nNOS). In our first Aim we will determine if Ca2+-permeable AMPA receptors can activate successive regulatory phosphorylations of nNOS by Akt and CaMKII, previously shown by us (Rameau et al., 2007) to be induced by the NMDAR. We will study scaffolding structures and biochemical pathways Akt and CaMKII that contribute to nNOS control by Ca2+-permeable AMPARs. GluR2 when modified by RNA editing dominantly blocks the Ca2+ conductance of AMPAR channels. In contrast, unedited GluR2 is highly toxic through its Ca2+ permeability and constitutive trafficking (Mahajan and Ziff, 2007). Recently we have found that NMDAR activity can degrade the GluR2 pre mRNA editing enzyme, ADAR2. Our second Aim is to analyze the pathological, NMDAR-dependent proteolytic cleavage of ADAR2, the production of unedited GluR2 as RNA editing activity declines, and the mechanisms by which Ca2+-permeable AMPARs including the unedited GluR2 can kill neurons. To limit Ca2+-toxicity, cells have evolved mechanisms for restricting synaptic AMPAR Ca2+ currents in which Ca2+-impermeable AMPA receptors replace Ca2+-impermeable ones at the synapse. In preliminary studies, we have found that release of Ca2+ from ER stores cooperates with CaMKII to stimulate trafficking of GluR2 from the endoplasmic reticulum to the plasma membrane. In our third Aim, we will study the Ca2+-dependent trafficking of GluR2 from the ER, and distinguish if its control relies on the assembly of GluR2 into tetramers or release of GluR2 from ER retention. We will determine the domains of GluR2 that respond to Ca2+ and the roles of intracellular Ca2+ levels, CaMKII, PICK1 in the export mechanism, including the roles of PICK1-CaMKII complexes. These studies will provide a comprehensive analysis of physiologic and pathologic pathways controlled by Ca2+-permeable AMPARs and mechanisms for regulating Ca2+-permeable AMPAR function.